JPS6136297A - Manufacture of carboxyalkyldipeptide - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION A typical method for preparing carboxyalkyl dipeptides and derivatives thereof is described in U.S. Pat. No. 4,374,829.
Disclosed in the issue. Although these methods are satisfactory, they are not commercially attractive. This is because the protecting groups used are expensive, relatively unstable and require additional separate reactions for stepwise removal. Examples of these prior art protecting groups are N-formyl, N-t-butoxy! These are compounds such as levonyl, N-carbobenzyloxy, phthaloyl, and succinoyl. Furthermore, the use of such protecting groups requires more steps and, as a result, longer and more expensive processing conditions.

It has been found that the carboxyalkyl dipeptides disclosed in US Pat. No. 4,374,829 can be obtained at lower cost and under shorter reaction conditions using the improved process of the present invention. The method of the present invention uses less expensive and more stable protecting groups than the typical protecting groups used in prior art methods, which are easily removed at the same time as obtaining the desired final product. , which eliminates the need for a separate time-consuming and costly reaction.

The protecting group used in the method of the invention is an α-fufulated acyl group represented by the following general formula.

-C-C- F A where: 2 is unsubstituted or substituted Cl-C1
2 alkyl, the substituent is halogen (F, Br,
α, I), lower c, -c8 alkoxy, aryl is C
6 or C10 aryloxy, di-lower C, C8 alkyl, lower C, -C, alkylthio, C6 or C
10 arylthio; unsubstituted or substituted C6 or C1o aryl whose substituents are lower C, -C8 narkyl, lower C1-C8 alkoxy, or halogen (FlBr, α, ) is a heterocyclic group up to C7 containing one OSN or S heteroatom.

A preferred protecting group is trifluoroacetyl (TFA).

It is disclosed in U.S. Pat. No. 4,374,829, which is hereby incorporated by reference.

These carboxyalkyl dipeptide compounds are represented by the formula: R"OR7I where R and R6 are the same or different, hydroxy, lower C, -C8 alkoxy lower C, -C8 alkenoxy, di-lower C, -C8 alkylamino lower c, -c8 alkoxy (dimethylaminoethoxy), acylamino lower C, -C8 alkoxy, (acetylaminoethoxy), acyloxy lower C, -C8 alkoxy, (pivaloyloxymethoxy), Aryloxy (e.g. phenoxy), aryl lower CI c.

alkoxy (e.g. benzyloxy), substituted aryloxy or substituted aryl lower c, -C8 alkoxy is substituted with methyl, halogen or methoxy), lower C,C8 alkylamino, di(lower C,-C8 alkyl) amino, hydroxyamino, aryl lower C,-C8 alkylamino (e.g. benzylamino); R1 is hydrogen, hydrocarbons of 1 to 20 carbon atoms (branched and unsaturated (e.g. C3CIQ cycloalkyl substituted lower CI-CB alkyl (substituent is halogen, hydroxy, lower C1-CB alkoxy; aryloxy (e.g. phenynoxy); amino; di(lower c, -c8 alkyl) Acylamino (e.g. acetamide and benzamide); Arylamino; Guanidino; Imidazolyl; Indolyl; Mercapto; Lower C,-C8 alkylthio; Arylthio (e.g. phenylthio); Carboxy or carboxamide; e.g. phenyl or naphthyl); substituted aryl (
For example, if the substituent is lower c, -, 'c8 alkyl, lower C
1-C8 alkoxy or halogen), aryl lower C,-C.8 alkyl, allyl lower C2-
C8 alkenyl, heteroaryl lower C, -C, alkyl 191) ^^ or heteroaryl lower C2'-C8 alkenyl (e.g. pencil, styryl or indolyl ethyl), substituted aryl lower C, -C alkyl, substituted aryl lower C, -C8 alkenyl, substituted heteroaryl lower C2-C8 alkyl, or substituted heteroaryl lower C2-C8 alkenyl, C whose substituent is halogen, dihalogen, lower C1-C8 alkyl, Hydroxy, lower c, -C8 alkoxy, amino, aminomethyl, acylamino (acetylamino or benzoylamino, diC lower c, -C8 alkyl) amino, lower c, -C8 alkylamino, carboxyl, halogen lower CI C8 alkyl, cyano or sulfonamide); aryl lower C, -C8 alkyl or to, (22) teroaryl lower c, -c8 (the alkyl moiety is substituted by amino or acylamino (acetylamino or benzoylamino)); R7 are the same or different and are hydrogen or lower c, -c8 alkyl; R3 is aminomethylphenyl lower C, -c8 alkyl; amino lower c, -c8 alkyl; guanidino lower C, -08 Alkyl; imidazolyl lower C, -C, alkyl; indolyl lower C, -C8 alkyl; mercapto lower C, -C8 alkyl; monoalkylamino lower alkyl (this alkyl group is c, -C8); R4 is hydrogen or lower C, -C8 alkyl; R5 hydrogen,
Lower C, -C8 alkyl, phenyl, phenyl lower C,
-C8 alkyl, hydroxyphenyl lower c1 -C8 alkyl, hydroxy lower C1-08 alkyl, amino lower c, -C8 alkyl, guanidino lower C1-C8 alkyl, imidazolyl lower ct C8 alkyl, indolyl lower C, -C8 alkyl, mercapto lower C, -C8 alkyl or lower c, -c8 alkylthio lower C1-C8 alkyl; or R4 and R5 are joined to form an alkylene bridge of 2 to 4 carbon atoms, 2 to 3 carbon atoms and a sulfur atom; fullkylene bridge, an alkylene bridge consisting of 3 to 4 carbons containing one double bond, or as above hydroxy, lower C,'-C8 alkoxy, lower c, -c8 alkyl or di-lower C , -C, an alkylene bridge substituted with alkyl.

In the groups 2 and R-R', "acyl" represents a group derived from an organic acid up to C6, and "aryl" and the prefix LS ar // represent an aromatic group of C6-C1°. represents a ring, and 1hetero “represents one of the O, N, or S heteroatoms.

By applying the method of the present invention, alanine, arginine, asparagine, aspartic acid, cysteine, cystine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, ornithine, phenylalanine, proline, serine, threonine,
Compounds of formula 1 or similar compounds can be obtained having peptides containing amino acids that are members selected from the group consisting of tryptophan, tyrosine, and/or valine.

Among the compounds of formula 1 above, compounds of particular interest are:

(a) N-(1-carboxy-3-phenylpropyl)-L-lysyl-L-proline and its acid salt; (b) N-(1-carboxy-5-aminopentyl)-L-alanyl-L -Proline; (c) N-α-
(1-carboxy-3-phenylpropyl)-L-lysyl-L-proline; (d) N-α-(1-ethoxycarbonyl-3-phenylpropyl)-L-lysyl-L
-proline; (e) N-α-[1-carboxy-3-(3-indolyl)propyl)-L-lysyl-L-proline; (f) N-α-[1-carboxy-3-(4 -chlorophenyl)propyl]-L-lysyl-proline; (gl N-α-[-1-carboxy-2-phenylthioethyl]-L-lysyl-L-proline (h) N-α-[1- hydroxy-3-(4-chlorophenyl)propyl]-L-lysyl-trans-4-
Methoxy-L-proline; (i) N-α-[1-ruboxy-5-aminopentyl]-L-lysyl-proline; (j) N-α-(1-ruboxy-3-phenylpropyl )-L-ornithyle-L-proline; (klN-(:1-ethoxycarbonyl-3-(4-imidazolyl)propyl)-L-alanyl-proline; 4-imidazolyl)propyl)-L-lysyl-L-proline; (-N-α-[1(Sl-carboxy-3-phenylpropyl]-L-lysyl-L-proline; and (n) N-α- (1(S)-ethoxycarbonyl-
3-phenylpropyl]-L-lysyl-L-proline.

In general, the methods of the invention involve preparing a peptide or amino acid that is protected with the protecting group of the method of the invention; coupling the protected peptide or amino acid with another peptide or amino acid to create a protected peptide or amino acid; When you get a dipeptide;
reductively alkylating the protected dipeptide with a keto acid (or ester, or amide, or hydroxamide thereof); and simultaneously hydrolyzing the reductively alkylated peptide to remove the protecting group therefrom; and obtaining the desired carboxydipeptide compound.

The method of the invention is further illustrated by the following reaction scheme.

Reaction Schematic % Formula % As shown in this reaction scheme, L-lysine (L) is treated with an ester of trifluoroacetic acid (TFA ester) to produce TFA-L-lysine 2; i.e., Nε-trifluoroacetyl - Obtain L-lysine. When TFA-L-lysine is then treated with phosgene and combined with L-proline, a protected diptide compound; -N-carboxyanhydride C compound J, TFA-lysine NCA). The protected dipeptide was reductively alkylated with an appropriate α-ketoester to obtain Nε-trifluoroacetyl-Nα-[(S
)-α-carboxyalkyl]-L-lysyl-L-proline (6) is obtained.

Hydrolysis of 6 in the presence of a base gives Nα-carboxyalkyl-L-lysyl-proline dipeptide compound ①.

The method of the invention is further demonstrated in the following examples, in which all parts and percentages are by weight and all temperatures are in °C unless otherwise indicated.

Example I Nε-trifluoroacetyl-L-lysyl-L-proline (5) L-proline (10,1 g), potassium hydroxide (4,8
9g), potassium carbonate (17.3g), tetrahydrofuran (THF) (250mA) and water (325rnl)
A mixture was prepared and cooled to 0°C. To this stirred mixture was added Nε-trifluoroacetyl-L-lysine-N-carboxyanhydride (3
) (22,3,!9) solution was added. The pn of this mixture was adjusted to 5.8 with sulfuric acid. By high pressure liquid chromatography (HPLC) assay, Nε-trifluoroacetyl-L-lysyl-monoproline (TFA-
Yield of lysylproline) (5) is 26.6g (94%)
Met. This is an amorphous product whose N
, was confirmed as a salt of N-dicyclohexylamine.

Example 2 Nε-trifluoroacetyl-L-lysyl-pH5,8
An aqueous solution containing Nε-trifluoroacetyl-di-lysiluproline (5) (8 g) and potassium sulfate (6.2 g) was frozen and lyophilized under vacuum. The solid residue was stirred with 2-propanol (50d) to drive off undissolved potassium sulfate. N,N-dicyclohexylamine (4,27,!i+) was added to Solution F. The solution was seeded and crystallized at 5° C. for 3 days. This crystalline product was treated with cold 2-propanol (15d).
and drying under vacuum to give compound 9.139.

M, P, 147-149°C I dec,).

Calculated value (-C,, H2oF3N304- C1□H2,
(as N): C, 57,67; H, 8, 32: F, 10.95: N, 10.76° Actual value: C, 56, 43: H, 8, 95: F, 10.7
8; N110.44.

The purity as determined by HPLC was as high as 99%.

Example 3 Nα-[1-(S)-Ethoxycarbonyl-3-phenylpropyl-Nε-trifluoroacetyl-L-lysyl-L-proline (6) Solution of TFA-lysylproline (73,6 g) in water and THF vacuum 200#+7 below
! It was concentrated to 1-Butanol (200m) was added and the mixture was heated to 2007! It was concentrated to This concentration operation was repeated twice. The salts were then filtered off and the salt cake was washed with ethanol (400-). -Add another ethanol (1 t), Raney nickel (7 t) to the alcoholic liquid
, 5 teaspoons), powdered 3-layer molecular sieves (200 μl), and ethyl 2-oxo-4-phenylbutylide (68,29) were added. The slurry was heated to 40 psi (2,81 Kg/i) of hydrogen. below 25℃
It was stirred with When the hydrogen was absorbed, the solids were washed separately with ethanol (5007ml).The combined solution F was concentrated to a low volume and water (500ml) was added followed by methylene chloride (500ml).The pH The pH of the aqueous layer was adjusted to 9.2 with sodium hydroxide, and the methylene chloride layer was discarded.The aqueous layer was extracted twice or more with CH2α2, and these extracts were also discarded.The pH of the aqueous layer was adjusted to 4.6 with hydrochloric acid. and extracted with methylene chloride. The extract was dried over sodium sulfate and then concentrated to an oil. The oil was dissolved in methyl t-butyl ether (650 mg) and the desired product crystallized over several hours. did.

This slurry was added very slowly to cyclohexane (24
0me), stirred for several hours, and filtered with N
α-[1-(S)-Ethoxycarbonyl-3-phenylpropyl)-Nε-trifluoroacetyl-L-lysyl-proline (6), 54.8 g, m, p, 64 70°C
gave. A second crop of product (13.9 g) was obtained from the concentrate. 10g from methyl t-butyl ether
Recrystallization of 9.1,! i', m, p, 74-
It gave a recovery of 77°C which was shown to be the pure isomer by HPLC.

Calculated value (as C2, H34F3N306): C, 56
,70:H,6,47:N,7.94° Actual value: C156,73;H,6,54;N,7.75
゜25℃ Specific optical rotation [α] D = -24.7° (0.1NHα/CH
30H) 25℃ [α]4o5nr+1 knee 53,8°(0,I NHC
l! /CH30H) Example 4 Nα-[1-(S)-ruboxy-3-phenyNα-[
1-(S)-Ethoxycarbonyl-3-phenylpropyl]-Nε-trifluoroacetyl-L-lysyl-L-
Proline (6) (log) is converted to sodium hydroxide (6,0
5 g) was dissolved in water (60 d) and stirred for about 3 hours.

The pH was adjusted to 2.0 with concentrated hydrochloric acid and treated with a strongly acidic (sulfonic acid type) ion exchange resin in an acid cycle. Once the product was adsorbed, the column was washed with water to remove salts. The product was eluted from the ion exchange using an aqueous 4 tipyricine solution. Combine the eluate and add approximately 100 rne
The pH was adjusted to 562 with hydrochloric acid. This solution was concentrated to about 16 g, and the concentrated liquid was converted into ethanol (140 m
diluted with l). The solution was concentrated to about 22 I and rediluted with ethanol to a concentration of about 8-12 parts water (80-120 parts water per rnl! of solution). The product slowly crystallized. It was then filtered, washed with 9:1 ethanol-water, and dried to give Nα-[1-(S)-lypoxy-3-phenylpropyl]-L-lysyl-monoproline dihydrate. 7.6-7.9g (90-94%)
gave.

Percent water measured with Karl Fischer reagent =
8.2 Coefficient optical rotation, [α]. = -26,8° (0 in CH30H
．． IN hydrochloric acid).

Claims (1)

[Claims] 1. A method for producing a carboxyalkyl dipeptide compound, which comprises: (a) Formula: ▲ Numerical formula, chemical formula, table, etc. ▼ A protected α-fluorinated acyl group represented by ¥A¥ protecting the peptide with a group [wherein Z is halogen, unsubstituted or substituted C_1-C_2 alkyl, the substituent is halogen, lower C_1-C_8 alkoxy, aryl is C_6 or C_1_0 aryloxy, di(lower C_1-C_8 alkyl), lower C_1-C_8 alkylthio, aryl C_6 or C_1_0 arylthio; unsubstituted or substituted C_6 or C_1_0 aryl; and aryl whose substituent is lower C_1-C_8 alkyl, lower C_1-C_8 alkoxy, or halogen; a heterocyclic group up to C_7 containing one O, N, or S heteroatom] (b) coupling the protected peptide or amino acid with another peptide or amino acid to obtain a protected dipeptide; (c) reductively alkylating the protected dipeptide with a keto acid or ketoester; and (d) hydrolyzing the reductively alkylated dipeptide and simultaneously removing the protecting group therefrom to recover the carboxyalkyl dipeptide compound. 2. The method according to claim 1, wherein the protecting group is trifluoroacetyl. 3. Peptides are included in amino acids such as alanine, arginine, asparagine, aspartic acid, cysteine, cystine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, ornithine, phenylalanine, proline, serine, and threonine. , tripliphan, tyrosine, and/or valine. 4. The method according to claim 1, wherein the carboxyalkyl dipeptide compound has the general formula: ▲There are mathematical formulas, chemical formulas, tables, etc.▼. [In the formula, R and R^6 are the same or different, and include hydroxy, lower C_1-C_8 alkoxy; lower C_2-C_8 alkenoxy; di(lower C_1-C_8 alkyl)amino lower C_1-C_8 alkoxy; lower acylamino C_1-C_8 alkoxy; acyloxy lower C_1-C_8 alkoxy; aryloxy; aryl lower C_1-C_8 alkoxy; substituted aryloxy or substituted aryl lower C_1-C_8 alkoxy, where the substituent is methyl, halogen, or methoxy ); amino; lower C_1-C_8 alkylamino; di(lower C_1-C_8 alkyl) amino; hydroxyamino, aryl lower C_1-C_8 alkylamino; R^1 is hydrogen; hydrocarbon of 1 to 20 carbon atoms (min. (including branched and unsaturated groups); C_3-C_1_0 cycloalkyl; substituted lower C_1-C_8 alkyl (where the substituents are halogen, hydroxy, lower C_1-C_8 alkoxy); aryloxy; amino; di(lower C_1 -C_8 alkyl) amino; acylamino; arylamino; guanidino; imidazolyl; indolyl; mercapto; lower C_1-C_8 alkylthio; arylthio; carboxy or carboxamide; carbo-lower C_1-C_8 alkoxy; C_1-C_8 alkyl, lower C_1-C_8 alkoxy or halogen); aryl lower C_1-C_8 alkyl; aryl lower C_2-C_8 alkenyl; heteroaryl lower C_1-C_8 alkyl or heteroaryl lower C_2-C_8 alkenyl; Aryl lower C_1-C_8 alkyl, substituted aryl lower C_2-C_8 alkenyl, substituted heteroaryl lower C_1-C_8 alkyl, or substituted heteroaryl lower C_2-C_8 alkenyl, (one or more of them) The substituent is halogen, dihalogen, lower C_1-C_8 alkyl, hydroxy, lower C_1-C_8 alkoxy, amino, aminomethyl, acylamino, di(lower C_1-C_8 alkyl)amino, lower C_1-C_8 alkylamino, carboxyl, lower halogen C_1-C_8 alkyl, cyano or sulfonamido); aryl lower C_1-C_8 alkyl or heteroaryl lower C_1-C_8 alkyl (the alkyl moiety is substituted by amino or acylamino); R^2 and R^ 7 are the same or different,
hydrogen or lower C_1-C_8 alkyl; R^3 is aminomethylphenyl lower C_1-C_8 alkyl; amino lower C_1-C_8 alkyl; guanidino lower C_1-C_8 alkyl; imidazolyl lower C_1-C_8 alkyl; indolyl lower C_1-C_8 alkyl ; mercapto lower C_1-C_8 alkyl; monoalkylamino lower alkyl (this alkyl group is C_1-C_8); R^4 is hydrogen or lower C_1-C_8 alkyl; R^5
is hydrogen; lower C_1-C_8 alkyl; phenyl; phenyl lower C_1-C_8 alkyl; hydroxyphenyl lower C_1-C_8 alkyl; hydroxy lower C_1-C_8 alkyl; amino lower C_1-C_8 alkyl; guanidino lower C_1-C_8 alkyl; imidazolyl lower C_1 -C_6 alkyl; indolyl lower C_1-C_8 alkyl; mercapto lower C_1-C_8 alkyl or lower C_1-C_8 alkylthio lower C_1-C_8 alkyl; or R^4 and R^5 combined to form 2 to 4 carbon atoms alkylene bridge of atoms; alkylene bridge of 2 to 3 carbon atoms and sulfur atoms; alkylene bridge of 3 to 4 carbon atoms with double bond; or hydroxy, lower C_1 as above -C_8 alkoxy, lower C_1-C_8 alkyl or di(lower C_1-C_8 alkyl) to form an alkylene bridge]. 5. The method of claim 4, wherein the carboxyalkyl dipeptide compound is a member selected from the group consisting of: {(a) N-(1-carboxy-3-phenylpropyl)-L-lysyl-L-proline and its acid salts; (b) N-(1-carboxy-5-aminopentyl)-L-alanyl- L-proline; (c) Nα-(1-carboxy-3-phenylpropyl)-L-lysyl-L-proline; (d) Nα-(1-ethoxycarbonyl-3-phenylpropyl)-L-lysyl-L -Proline; (e) Nα-[1-carboxy-3-(3-indolyl)propyl]-L-lysyl-L-proline; (f) Nα-[1-carboxy-3-(4-chlorophenyl)propyl] -L-lysyl-L-proline; (g) Nα-[-1-carboxy-2-phenylthioethyl]-L-lysyl-L-proline (h) Nα-[1-carboxy-3-(4-chlorophenyl) ) propyl]-L-lysyl-trans-4-methoxy-L-proline; (i) Nα-[1-carboxy-5-aminopentyl]-L-lysyl-L-proline; (j) Nα-(1- (k) N-[1-ethoxycarbonyl-3-(4-imidazolyl)propyl]-L-alanyl-L-proline; (l) N- [1-carboxy-3-(4-imidazolyl)propyl]-L-lysyl-L-proline; (m)Nα-[1(S)-carboxy-3-phenylpropyl]-L-lysyl-L-proline; and (n) Nα-[1(S)-ethoxycarbonyl-3-phenylpropyl]-L-lysyl-L-proline}. 6. A compound having the formula: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ [X in the above formula is an α-fluorinated acyl protecting group represented by the formula: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ and Z in the formula is C_1-C_1_2 alkyl that is not substituted or substituted with halogen, and the substituent is halogen, lower C_1-C_8 alkoxy, aryloxy (this aryl is C_6 or C_1_0) ), di-lower C_1-C_6 alkyl, lower C_1-C_8 alkylthio, aryl C_6 or C_1_0thio; unsubstituted or substituted aryl of C_6 or C_1_0, the substituent being Lower C_1-C_8 alkyl, lower C_1-C_8
aryl which can be alkoxy, or halogen; is a heterocyclic group up to C_7 with one O, N or S heteroatom]. 7. The compound according to claim 6, wherein X is the formula: ▲A mathematical formula, a chemical formula, a table, etc.▼. 8. A compound having the formula: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ [wherein X is an α-fluorinated acyl protecting group represented by the formula: ▲There are mathematical formulas, chemical formulas, tables, etc.▼, Z in the formula is unsubstituted or substituted C_1-C_1_2 alkyl, and the substituent is halogen, lower C_1-C_8 alkoxy, aryloxy (the aryl is C_6 or C_1_0), di Alkyl which can be lower C_1-C_8 alkyl, lower C_1 -C_8 alkylthio, aryl C_6 or C_1_0thio; unsubstituted or substituted aryl of C_6 or C_1_0, the substituent being lower C_1- Aryl, which can be C_8 alkyl, lower C_1 -C_8 alkoxy, or halogen; heterocyclic groups up to C_7 with one O, N or S heteroatom; and Y is unsubstituted or substituted C_1-C_1_2 alkyl whose substituents can be halogen, hydroxy, C_1-C_8 alkoxy; C_6-C_1_0 aryl which is unsubstituted or substituted; aryl where the substituents can be hydroxy, C_1-C_8 alkyl, C_1-C_8 alkoxy]. 9. The compound according to claim 8, wherein X is ▲a mathematical formula, a chemical formula, a table, etc.▼.